Production of Lentiviral Vector Expressing MicroRNA-148b

Document Type : Original Article (s)

Authors

1 PhD Candidate, Biotechnology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

2 Professor, Biotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

3 Assistant Professor, Medical Genetics Research Center AND Department of Medical Genetics, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

Abstract

Background: Micro (mi)RNAs are non-coding endogenous RNAs which regulate gene expression by hybridization to specific binding sites in target mRNA sequences. Since several miRNAs are involved in proliferation and differentiation, miRNA-based therapies could be promising approach in regenerative medicine. Among different vehicles, lentiviral vector system is suitable for miRNA delivery. Besides, it is shown that miRNA-148b is involved in osteogenic differentiation. In this study, designing and cloning of miR-148b to lentiviral vector were investigated.Methods: We introduced miRNA-148b-3p/-5p into lentiviral vector through cloning producers. The sequences of lentiviral vectors carrying miRNA-148b were checked via analytical digestion as well as Sanger DNA sequencing. In the following, produced lentiviral vectors were used for mesenchymal stem cells transduction.Findings: Designed miR-148b-3p/-5p successfully cloned to the shuttle. Correctness and absence of any unintended mutations of lentiviral shuttle carrying miRNA-148b3p/-5p were confirmed followed by lentiviral production. Expression of enhanced green fluorescent protein (eGFP) demonstrated high efficiency of transfection as well as transduction.Conclusion: Viral vectors constructed in this study could be used for investigation of osteogenesis.

Keywords

References

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Journal of Isfahan Medical School
Volume 35, Issue 434 - Serial Number 434
May and June 2017
Pages 701-706

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History

  • Receive Date: 01 March 2017
  • Revise Date: 01 November 2024
  • Accept Date: 06 April 2022

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